用PGA支架体外构建人口腔黏膜固有层的实验研究

Experimental study on in vitro lamina propria engineering using oral fibroblast and polyglycolic acids

目的:探讨利用可吸收生物材料聚羟基乙酸(polyglycolicacids,PGA)和口腔黏膜成纤维细胞(oralfibroblast,OFC)在体外构建组织工程化口腔黏膜固有层的可能性。方法:取细胞经体外培养,扩增至第3代,与PGA混合培养,形成细胞—生物材料复合物,每2d换液一次。动态观察细胞形态和粘附生长状况,于体外培养1周左右,取组织作电镜观察、组织学和RT-PCR检测。结果:复合物体外培养6d,OFC粘附在生物支架上,沿PGA纤维纵轴方向向两极伸展,细胞分泌基质并形成拉网状结构。HE染色和Masson染色均提示胶原纤维形成,RT-PCR显示胶原成分主要为Ⅰ型胶原。结论:应用PGA支架可以在体外成功构建口腔黏膜固有层组织。

PURPOSE: This study investigated the feasibility of lamina propria engineering in vitro using expanded oral fibroblast(OFC) and Polyglycolic Acids (PGA). METHODS: OFC were isolated by tissue explant method and expanded in vitro. OFC (20×106) of 3rd passage were collected and then seeded onto PGA unwoven fibers to form a cell-scaffold. The constructs were cultured in DMEM +10% FBS. The cell-scaffold constructs were observed continuously by microscope. Small fragments were harvested at 1 week for electromicroscope, histological and RT-PCR analysis. RESULTS: At the sixth day, a neo-lamina propria was formed. HE and Masson stain revealed the formation of collagen fibers. RT-PCR revealed the new forming collagen was mainly typeⅠcollagen. CONCLUSION: lamina propria tissue is possible to engineer in vitro using oral fibroblast and polyglycolic acids. At this basis, we can construct bi-layer tissue engineering oral mucosa in the further research.